PCR conditions:
Pre heated lid: 111°C
Denaturation: 94°C for 10min
Cycle (33×)
Denaturation: 94°C for 30s
Annealing: 60°C for 30s
Extenstion: 72°C for 2min
Final extension: 72°C for 10min
Store infinite at 4°C
Analyse PCR samples on a 1% agarosegel
7 out of 9 for PhybB-RBS-cI-LVA-DT have the correct size and 5 out of 9 for PhybB-RBS-LasR-LVA-DT have the correct size!
Send them monday for sequencing